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Abstract
In type 1 diabetes, the immune system destroys pancreatic beta cells in an autoimmune condition. To overcome this disease, a specific monoclonal antibody that binds to pancreatic beta cells could be used for targeted immunotherapy. Protein tyrosine phosphatase receptor N (PTPRN) is one of the important surface antigen candidates. Due to its high sequence homology among mammals, so far, no single-chain monoclonal antibody has been produced against this receptor. In this study, we developed a novel single-chain variable fragment (scFv) against the PTPRN extracellular domain. To this aim, ostrich species was used as a host is far phylogenetically birds from mammals to construct a phage display library for the first time. An ostrich-derived scfv phage display library was prepared and biopanning steps were done to enrich and screen for isolating the best anti-PTPRN binders. An scFv with appropriate affinity and specificity to the PTPRN extracellular domain was selected and characterized by ELISA, western blotting, and flow cytometry. The anti-PTPRN scFv developed in this study could be introduced as an effective tool that can pave the way for the creation of antibody-based targeting systems in cooperation with the detection and therapy of type I diabetes.
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Details
1 Tarbiat Modares University, Department of Genetics, Faculty of Biological Sciences, Tehran, Iran (GRID:grid.412266.5) (ISNI:0000 0001 1781 3962)
2 Pasteur Institute of Iran, National Cell Bank of Iran, Tehran, Iran (GRID:grid.420169.8) (ISNI:0000 0000 9562 2611)
3 ACECR, Department of Regenerative Medicine, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, Tehran, Iran (GRID:grid.419336.a) (ISNI:0000 0004 0612 4397); ACECR, Advanced Therapy Medicinal Product Technology Development Center (ATMP-TDC), Royan Institute for Stem Cell Biology and Technology, Tehran, Iran (GRID:grid.419336.a) (ISNI:0000 0004 0612 4397)